Allergy Asthma Immunol Res.  2013 Nov;5(6):377-382. 10.4168/aair.2013.5.6.377.

IL-13 R110Q, a Naturally Occurring IL-13 Polymorphism, Confers Enhanced Functional Activity in Cultured Human Bronchial Smooth Muscle Cells

• He YF
• Hua L
• Bao YX
• Liu QH
• Chu Y
• Fang DZ

• KMID: 2260281
• DOI: http://doi.org/10.4168/aair.2013.5.6.377

Abstract

PURPOSE
Interleukin (IL)-13, a Th2-type cytokine, plays a pivotal role in the pathogenesis of asthma through its direct effects on airway smooth muscles. A naturally occurring IL-13 polymorphism, R110Q, is strongly associated with increased total serum IgE levels and asthma. In the present study, we aimed to determine whether the IL-13 R110Q variant would display different biochemical properties or altered functions in comparison with wild-type (WT) IL-13 in cultured human bronchial smooth muscle cells (hBSMCs).
METHODS
Culture supernatants and cell proteins were collected from cultured hBSMCs that were treated with 50 ng/mL IL-13 or IL-13 R110Q for 24 hours. Eotaxin released into hBSMC culture medium was determined by ELISA. The expression levels of the high-affinity IgE receptor (FcepsilonRI) alpha-chain, smooth muscle-specific actin alpha chain (alpha-SMA), smooth muscle myosin heavy chain (SmMHC), and calreticulin in the cells were measured on Western blots.
RESULTS
Compared with WT IL-13, treatment with the IL-13 R110Q variant resulted in a significant increase in eotaxin release as well as significant, although modest, increases in the expression levels of alpha-SMA, SmMHC, calreticulin, and FcepsilonRI alpha-chain.
CONCLUSIONS
The results of the present study suggenst that the IL-13 R110Q variant may enhance enhanced functional activities in hBSMCs.

Keyword

IL-13; polymorphisms; bronchial smooth muscle cells; functional activity

MeSH Terms

Actins
Asthma
Calreticulin
Enzyme-Linked Immunosorbent Assay
Humans
Immunoglobulin E
Interleukin-13
Interleukins
Muscle, Smooth
Myocytes, Smooth Muscle
Myosin Heavy Chains
Proteins
Actins
Calreticulin
Immunoglobulin E
Interleukin-13
Interleukins
Myosin Heavy Chains
Proteins

Figure

• 

  Fig. 1 hBSMCs were treated with the indicated concentrations of WT IL-13 or the IL-13 variant (1, 10, 50, and 100 ng/mL) for 24 h. Following treatment, culture supernatants were collected, and the eotaxin concentration was measured by ELISA. Data are presented as means±SD from three or more samples.

• 

  Fig. 2 Cellular proteins were obtained from hBSMCs treated with WT IL-13 or the IL-13 R110Q variant (50 ng/mL each) for 24 h and used for Western blot analysis of (A) α-SMC and (B) SmMHC proteins. Data are presented as means±SD from three or more samples.

• 

  Fig. 3 Cellular proteins were collected from hBSMCs treated with WT IL-13 or the IL-13 R110Q variant (50 ng/mL each) for 24 h and used for Western blot analysis of (A) FcεRI α-chain and (B) calreticulin proteins. Data are presented as means±SD from three or more samples.

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